目的 研究非离子型的diazeniumdiolate类一氧化氮供体引起肝癌细胞凋亡的分子机制。方法 利用免疫印迹、免疫沉淀、凝胶阻滞实验研究一氧化氮供体处理Hep3B肝癌细胞后,丝裂原激活的蛋白激酶、AP-1的激活以及和Hep3B肝癌细胞凋亡的关系。结果 一氧化氮可引起细胞外信号调节蛋白激酶、c-jun N末端激酶和p38激酶的激活,特别是细胞外信号调节的蛋白激酶的持续激活,其中细胞外信号调节的蛋白激酶和c-jun N末端激酶的特异的阻断剂U0126和JNK抑制剂Ⅱ可阻断AP-1的激活和Hep3B细胞的凋亡,而p38激酶的阻断剂SB203580不能阻断AP-1的激活和Hep3B肝癌细胞的凋亡。结论 一氧化氮通过激活细胞外信号调节蛋白激酶、c-jun N末端激酶,进而激活AP-1而引起Hep3B肝癌细胞的凋亡。

Objective To evaluate the underlying mechanisms of Hep 3B cell apoptosis induced by JS-K, a non-ionic diazeniumdio late derivative of nitric oxide (NO) donor.Methods The activities of mitogen-activated protein kinases (MAPKs), active protein 1(AP-1) and apoptosis were studied with Western blotting, immonoprecipatation, and electrophoretic mobility shift assay (EMSA) in Hep 3B cell line.Results NO induced activation of extracellular-signal-regulated protein kinase (ERK), c-Jun N-terminal kinase (JNK) and p38 kinase, especially ERK persistent activation, and also induced activation of AP-1.AP-1 activation and apoptosis of Hep 3B cell line was partially abolished by U0126 and JNK inhibitor II, inhibitors of ERK and JNK, but not by SB203580, a inhibitor of p38 kinase, indicating that ERK and JNK activation involved the AP-1 activation and apoptosis induced by JS-K in Hep 3B cell line.Conclusion JS-K induces Hep 3B cell line apoptosis through ERK and JNK activation, which further activated AP-1.